COPPER AND ZINC LEVELS I N SERUM FROM

HUMAN PATIENTS W I T H SARCOMAS

G. L. FISHER,PHD,* V. S. BYERS,
PHD,+ng. SHIFRINE, PHD,*A N D
A. S. LEVIN,MD*

Serum copper levels (SCL) a n d serum zinc levels (SZL) were evaluated i n 19
patients with sarcomas, 12 of which were osteosarcomas a t various stages. Patients with primary or metastatic osteosarcoma h a d elevated SCL, whereas
amputated osteosarcoma patients who were clinically tumor-free h a d nearly
normal SCL. Patients with primary osteosarcoma h a d elevated SZL, those with
metastases h a d depressed zinc levels, a n d amputated patients who were clinically
tumor-free had nearly normal SZL. T h u s , the ratio of SCL:SZL i n metastatic
osteosarcoma patients is higher than in patients with primary osteosarcoma. SCL
a n d SZL are compared to clinical histories for selected patients. Patients with
the more advanced disease a n d poorest prognoses had the most elevated SCL
a n d highest SCL:SZL ratios. I t appears that the determination of SCL a n d SZL
i n osteosarcoma patients may be of value i n prognosis a n d therapy evaluation;
furthermore, the ratio of SCL:SZL may be useful i n discriminating between
patients with primary a n d metastatic osteosarcoma.

Cancer 37:356-363. 1976.

LTHOUGH IT IS WELL K N O W N THAT THE ELE-

ments copper and zinc are biologically

essential for normal development, growth, and
function, very little is known about their
clinical significance in malignant diseases or
their value in diagnosis ant1 prognosis. Our
report describes changes in serum copper level
(SCL) and serum zinc level (SZL) in human
patients with osteogenic and other sarcomas.
Serum copper levels, which correlate with
From the Radiology Laboratory, School of Veterinary
Medicine, University of California at Davis, CA.
Suppotted by the U S . Atomic Energy Commission.
* Postgraduate Research Chemist, Radiobiology Laboratory, School of Veterinary Medicine, University of
California, Davis.
t Associate Research Immunologist, Department of
Medicine, University of California, San Francisco.
t Adjunct Professor of Immunology, Radiobiology
Laboratory, School of Veterinary Medicine, University
of California, Davis.
9 Adjunct Assistant Professor of Immunology in
Dermatology, IJnivcrsity of California Mcdical Center,
San Francisco; and Chief, Division of Immunology,
Western Laboratorics, Oakland, California.
Address for reprints: G. L. Fisher, PhD, Radiobiology Laboratory, University of California, Davis,
CA 95616.
The authors thank Dr. J . 0. Johnston, Department
of Orthopedics, Kaiser Hospital, Oakland, California,
for patient management and serum samples, and
Ellen K. Hato for editorial assistance.
Received for publication January 20, 1975.

the serum enzyme ceruloplasmin, have been

reported to be elevated in neoplastic disorders.
Warren et al.24 reported a close relation between Huctuating patterns of elevated serum
copper and the prognosis of active Hodgkins
disease. Similar results were recently reported
by Tessmer et al.,*3 who concluded that serum
copper levels are valuable in determining and
evaluating Hodgkins disease.
Hrgovcic et a1.10 showed that serum copper
reflects relapse or remission in cases of leukemia, thus indicating the efficacy of treatment. They reported elevated SCL in the active disease, decrease with response to therapy,
normal values during remission, and elevated
SCL in relapse. Other clinical tern (SGOT,
calcium, glucose, BUN, LDH) did not correlate well with serum copper analyses, underlining the value of SCL as an independent
index of disease prognosis and therapy evaluation in human patients with lympllomas and
acute leukemias. Similar results have been reported by Delves et al.,4 by Illicin,ll and by
Tessmer et al.z2
Fisher5 observed significant changes in serum
copper levels of osteosarcomatous beagles with
relatively high body burdens of 90Sr and n2sRa
and in dogs with spontaneous osteosarcoma.
T h e degree of SCL elevation appeared to be
related to disease activity, with return to nor-

356

No. I

SERUM
COPPERAND ZINCIN SARCOMA
PATIENTS

ma1 SCLs after amputation of affected limbs

in dogs with osteosarcoma of the appendicular
skeleton.
In a plot study on the effects of chronic
Wo-irradiation on beagles, Mac Millan et aL1*
observed significantly lower SCLs in irradiated dogs than in controls. These results are
of interest in light of the report by Hrgovcic
et al.1" that radiotherapy has led to decreases
in the elevated SCL's of human patients with
Hodgkin's disease.
As early as 1958, Addink and Frank1 reported that, unless neoplasms develop in tissues relatively rich in zinc (e.g., bone, lung),
whole blood and serum zinc levels in human
patients with such neoplasms are generally
subnormal, with favorable medical progress
accompanied by increase toward normal
values. Szmigielski and Litwin's21 report that
below-normal values of zinc in blood granulocytes can be used as a test in the diagnosis
of neoplastic disease has not been confirmed.
T h e role of zinc in cancer growth has been
investigated by many researchers. Direct evidence of a relation between dietary zinc levels
and tumor growth is shown by the work of
DeWys et aL,5 in which low-zinc diets retarded
the growth of Walker 256 carcinosarcoma in
rats and increased survival time after tumor
cell transplantation. Also, Chahovitch3 is reported to have shown that the administration
of zinc sulfate accelerated the growth of chemically induced sarcomas. Janes et al.13 demonstrated that the zinc concentration in human osteogenic sarcoma averaged 217%
greater than that in normal bone.
Investigating the role of ions in growth
regulation of cells in tissue culture, Rubin and
Kordils demonstrated that DNA synthesis that
had been inhibited in animal cells by ethylene
diamine tetraacetate (EDTA) was resumed
after the addition of zinc. Rubin also found
Rous sarcoma cells to be relatively resistant
to inhibition by EDTA and suggested that
malignant cells may have more free Zn++than
d o normal cells.
From a review of the literature, MikacDevic15 concluded that the contradictory data
on zinc levels in biologic specimens necessitate
further studies to define the usefulness of zinc
analyses in the diagnosis of cancer.
Osteosarcoma, a malignant lesion of osseous
origin, is a rare disease estimated to occur at
the rate of 1 in 100,000 persons.2 T h e five-year
survival rate of patients with osteosarcoma is
very low, estimated to be 5 to 15%,.8 T w o age

Fisher et al.

357

groups appear to be at greatest risk for development of osteosarcoma. T h e majority of

patients are 11 to 20 years of age; the second
group is 50 to 60 years of age. Most primary
osteosarcomas occur in the long bones of the
arms or legs and metastasize into the lung
within about 18 months after limb amputation. Currently there are no satisfactory ways
to follow the progress of the disease until the
secondary tumor can be detected ratliographically.
CLINICAL
MATERIAL
This study was performed on serum samples
from 19 sarcoma patients, 18 with bone involvement, referred to us by orthopedic surgeons in the San Francisco Bay Area. Twelve
of the bone tumors were osteosarcomas, at
stages ranging from initial biopsy and diagnosis through extensive metastatic disease to
death. Three patients had giant cell sarcomas
of bone, a tumor that is closely related to
osteosarcoma and that frequently develops
into osteosarcoma either spontaneously or in
response to radiation.19 T h e remaining four
patients included one with chonclrosarcoma of
bone, one with reticulum cell sarcoma of bone,
one with rhabdomyosarcoma, and one with
fibrosarcoma. T h e tumors were classified by
an orthopedic pathologist on the basis of clinical history, radiology, and histology following
open biopsy and/or amputation.
METHODS
Serum samples, obtained at the times indicated, were analyzed by conventional techniques on a Perkin-Elmer Model 306 Atomic
Absorption unit. Serum copper and zinc were
analyzed in 1:4serum:H,O dilution. All glassware was rinsed three times with dilute HNO,
in doubly distilled water, followed by three
rinsings with doubly distilled water. New
standards were prepared on the day of each
analysis. Serum was stored frozen in tightly
capped bottles, washed as described above.
Statistical analysis was performed by the
standard t-test when the I;-ratio of variances
indicated no significant differences in distribution. If the F-ratio was significant, then the
degrees of freedom for the t-test of samples
with unknown and unequal variances were
calculated using the Welch appr0ximation.~7
T h e comparisons reported for the osteosarcoma group include some multiple measure-

CANCER
J n n ti nry 19 7 6

358

VOl. 37

metastases had higher elevated SCLs than the

group with primaries. Furthermore, zinc levels
were lower in the group with metastases than
in the group with primary tumors only. T h e
average SCL:SZL ratio for individual patients
with primary tumors alone was 0.94 -+ 0.47
( 2 S.D.), whereas the average ratio for patients
with metastases was 2.28 -t- 0.43 (eS.D.); the
difference was highly significant (p < 0.001).
Patients who were clinically tumor-free after
amputation and while on therapy had nearly
normal SCLs, lower than either the primary
or metastasis groups. SZLs for the tumor-free
group were nearly normal, and lower, but not
significantly different, than the group with
primary tiimors only. T h e four patients with
primary sarcomas other than osteosarcoma had
SCLs the same as SCLs in patients with primary tumors only, and SZLs which were lower
but not statistically different from SZLs in
patients with primary osteosarcomas only. Patients with histories of osteosarcoma who were
clinically tumor-free did not have SCL:SZL
ratios which were significantly different than
the primary tumor group.
T h e results for individual patients with
regard to clinical observation and treatment
are presented in Table 4. T h e data indicate
that the extent and activity of disease are
associated with the degree of elevation of
SCLs. Clinical histories are compared with
SCL and SZL for exemplary patients and for
those for whom data were available at different times during the clinical course of the
disease. I t appeared from comparison of clinical observation with SCL values that elevation of SCL is greatest in patients with the
most active malignancies and the poorest
prognoses.

ments on the same patients at different times

during the course of the disease.

RESULTS
Results of analyses of serum from normal
patients are presented in Table 1. Normal
females had SCLs slightly higher (p
0.1)
than normal males. As noted by many authors,
we found SCLs to be elevated in women
taking oral contraceptives.. Data on these
women were removed from our normal
population since their mean SCL (220 pg/IOO
ml) was significantly different (p < 0.001) from
the controls. Our population mean for SCLs
in normals was the same as that reported by
Tessmer et a1.22 We also observed a significantly lower SZL in normal females than
males. Our overall mean SZL is similar to that
reported in Scientific Tables.6
Serum copper levels were found to be significantly elevated in the group of patients
with histories of sarcomas (Table 2). Also, the
group variances were found to be statistically
different. Serum zinc levels were elevated but
not statistically different, although the group
variances were different. Analyses of the serum
copper and zinc levels of osteosarcoma patients at different stages of the disease also
indicated significant differences (Table 3), despite the relatively small number of patients
in each group. These data are presented
graphically in Figs. 1-3. T h e values used in
each group are indicated in Table 4. I n comparing SCLs and SZLs in patients with primary osteosarcoma alone to those in patients
with both primary tumors and lung metastases or lung metastases alone, the group with

TABLE
1. Serum Cooper and Zinc Levels in Normal Human Controls
Normals

x*

(N)t

S.D.t

104
113
108

N.S.
-

1.71

(25)

14.04
12.59
13.96

1.24

()

(14)
(11)
(25)

14.19
9.08
14.41

2.44

N.S.
-

3.21

FWtiOS

cu
Male
Female
All
Zn
Male
Female
All

109

94
102

Mean in pg/100 ml.

Number of individuals.
Standard deviation.
F-ratio of variances for male-female comparison.
** Calculated t-statistic for male-fernale comparison.
+t Degrees of freedom.
t

Sig.

t(calc.)**

d.f.t+

Sig.

23

p -0.1
-

23

< 0.01
-

No. 1

SERUMCOPPER
AND ZINCIN SARCOMA
PATIENTS Fisher et al.

359

TARLE
2. Comparisons of SCLs and SZLs in Humans with Histories of Sarcomas and Normal Controls

x*

(N)

S.D>

F r a t io*

167
108

(19)
(25)

24.76
13.96

3.15

125
102

(19)
(25)

69.68
14.41

23.38

Sig.

l(calc.)**

d.f.+t

Sig.

< 0.01

9.26

27.5

p < < O 001

p <<0.001
-

1.42

19.3

N.S.

cu
Sarcoma
Normal

Zn
Sarcoma
Normal

* Mean in pg/100 ml.

t Number of individual values (with means for those patients with more than one observation).
t Standard deviation.

* F-ratio of variances for sarcoma-normal comparison.

** Calculated t-statistic for sarcoma-normal comparison.

Degrees of freedom calculated after Welchs method?.

tt

CASEHISTOR~ES
Patient 4 had the highest observed SCL and a
high SCL:SZL ratio. At the time of copper analysis,
the patient had a n osteosarcoma measuring 12
cm x 18 cm protruding from the posterior lip of
the right ilium and a solitary lung metastasis.
Rapid growth of the lung metastasis caused death
7 weeks later.
Patient 6 had osteosarcoma and a very high
SCL and a high SCL:SZL ratio. At the time the

serum sample was obtained the primary tumor

mass measured only 2 cm i n diameter, and a solitary lung metastasis was present. However, the
patient died 3 months later from extensive pulmonary metastases, indicating that the tumor was
very active.
Patient 9 had the highest observed SCL:SZL
ratio i n the presence of a markedly elevated SCL.
T h i s patient had large bilateral pulmonary metastases (20 cm diameter) a t the time the serum

TABLE
3 . Comparisons of SCLs and SZLs in Hunians with Osteosarcoma a t Different Stages of the Disease

cu

Comparison
of Types

x*

Primary
osteosarcoma,
nometastases
Metastatic
osteosarcoma

162

(5)

11.5

195

(6)

24.9

( N ) + S.D. FratiJ d.f.1 t(calc.)**

4.69

2.706i

x* S.D.
207

91.9

88

21.8

207

91 9

132

24 4

207

91 9

102

13.6

Zn
Cu : Zn
FrAti,,* d.f.b t(calc.)** ratio +t

17.7405

4.5

2.83*$

0.94
2.281t

~~~~~~~~~~~~~~~~

Primary
osteosarcoma,
no metastases
162
Amputated
osteosarcoma,
no demonstrable
tumor present
115
Primary
osteosarcoma,
no metastases
0ther sarcomas,
no metastases

(5)

11 5

(3)

25 4

162

(5)

11 5

166

(4)

11.4

4 87

1 02

3 74***

0 52

* Mean in pg/lOO ml.

t Number of individuals.

14 19

Calculated t-statistic relative to primary osteosarcoma.

individual serum copper to zinc ratios.

< 0.001.
< 0.05.
*** p < 0.01.

%*

65

p
p

1 34

0 94

0 91

45 6 6 s i

* F-ratio of variance relative to primary osteosarcoma.

* Degrees of freedom, corrected for significant variance ratio when appropriate.
**
t+ Average of

4 3

2 52

0 94
1.6455

CANCER
January 1976

360
230

2 I0

3 20

i2I
2 00

Val. 37

c
*

fcc

OL
50
NORMALS

PRIMARY OSTLO.
NO Y E T

METASTATIC

OSlEO

AUPUTArfO
OSTEO.NO

OTHER P R I M A R Y
SARCOMAS,

TUMOR PRESENT

NO MET

NORUIlS

PRIMARY O S T E O . Y E T P S T l T l C
NO M E T
OSTEO

AMPUTATED
O S T E O , NO

O T H E R PRIMLRY
SARCOMAS.

TUMOR PRESENT

NO YET

FIG. 3. Serum copper: zinc ratio. The mean value

is indicated by the horizontal bar.

FIG. 1. Serum copper levels. The mean value is

indicated by the horizontal bar.

tumor-free at present and will continue to b e
serially monitored.

was drawn and analyzed, and died within 2 months

after the sample was taken.

Patient 11 presented an unusual case in which

the initial diagnosis was an aneurysmal bone cyst
of the left humerus T h e patient received 2000R
to the lesion, which continued to spread locally
at a rapid rate. T w o months later, at the time of
biopsy, the tumor was reclassified as a giant cell
sarcoma, better described as a telangiectatic-type
osteogenic sarcoma. T h e first serum sample was
taken at this time. T h e second serum sample was
taken 3 months after the first at the time of
amputation of the affected humetus. At this time,
the tumor was reclassified as a n osteosarcoma. I n
spite of chemotherapy and immunotherapy this
patient died of pulmonary metastases 4 months
after amputation.

Patient 5 had a solitary pulmonary metastasis

in the left lung, which was resected. Serum taken
at the time of surgery had high copper levels and
a high SCL:SZL ratio. Serum taken 3 months later,
after resection, when the patient had been on
immunotherapy (transfer factor) and was clinically
tumor-free, had normal copper and zinc levels.
Patient 10 was similar to Patient 5 inasmuch as
SCL was high at the time the tumor mass was
present, and decreased to a normal level 4 months
after the mass was removed. T h e SCL was slightly
elevated 1 month later. This patient, now receiving
immunotherapy with transfer factor, is clinically

3 70

170)

50
NORMALS

PRIMARY

OSTEO. Y E l A S T A T l C

NO YET

OSTEO

A Y P U T I T L O OTHER PRIMARY
OSTE0,NO
SARCOMAS,
TUMOR PRESENT
NO M E T

FIG. 2. Serum zinc levels. The mean vaIue is indicated by the horizontal bar.

Patient 14 had had an osteosarcomatous tibia

amputated 8 months earlier. A serum sample was
obtained within 1 month after the patietlt presented with metastases. Results showed normal
SCL in the presence of depressed zinc. We have
n o explanation for the copper value; however, this
case was extremely unusual i n that one of the
metastases appeared in the proximal tibia, a location that would be expected to host a primary
tumor. However, 1 month after surgical removal
of the new tumors and during immunotherapy,
the patients SCL dropped below normal values,
implying that perhaps this patient had a normally
low serum copper level. Because of the unusual
nature of the metnstcises (located on long bones
in the epiphyseal region, which may habe been
new primaries) observed in this patient, the copper
and zinc values were not included in the comparison of osteosarcoma at different stages (Table 3).
Patient 15 demonstrated high SCL in two serum
samples but is not consideled in an osteosarcoma

SERUMCOPPER
AND ZINC IN SARCOMA
PATIENTS Fisher et al.

No. 1

36 1

TABLE
4. History of Tumor Patients and Serum Levels of Cu and Zn
Patient

Age
(yrs)

Sex

16

13

21

17

5a

20

5b

Same

13

12

18

18

10a

16

10b

Same

1oc

Same

1l a

llb

Same

12

30

13

38

14a

18

14b
I5a

15b

Same

17

Same

16

45

17

55

18

87

19

48

Clinical condition at time

of serum sample
Serum sample at time of amputation
of osteosarcoma 3 yr post diagnosis;
pulmonary metastasis present
Serum sample at time of amputation
of osteosarcoma; pulmonary
metastasis present
1 y r post amputation of osteosarcoma;
large pulmonary metastasis present
Large abdominal osteo~arcomapresent;
solitary pulmonary metastasis
Serum sample a t time of resection of
pulmonary metastasis 6 mo post
amputation of osteosarcoma
3 mo later; no clinical evidence
of disease
Osteosarcoma present; solitary
pulmonary metastasis
Serum sample at time of amputation
of osteosarcoma; no metastasis
Serum sample at time of amputation
of osteosarcoma 2 yr post diagnosis;
no metastasis
Serum sample a t time of biopsy prior
to amputation of small osteosarcoma;
no metastasis
Serum sample at time of amputation
of osteosarcoma; no metastasis
4 mo post amputation; no clinical
evidence of disease
5 mo post amputation; no clinical
evidence of disease
Serum sample a t time of biopsy of
giant cell sarcoma; no metastasis
3 mo later; serum sample a t time of
amputation of osteosarcoma;
no metastasis
Serum sample a t time of surgical
removal of giant cell sarcoma;
no metastasis
15 yr post amputation of giant cell
sarcoma; high cell-mediated immunity
against osteosarcoma; no clinical
signs of disease
8 mo post amputation of osteosarcoma;
new primary osteosarcoma present
1 mo later; no clinical signs of
disease; on immunotherapy
1 yr post amputation of osteosarcoma;
no clinical signs of disease; pregnant
1 mo later a t time of therapeutic
abortion
Serum sample at time of amputation
of fibrosarcoma; pulmonary metastasis;
cirrhosis of the liver
Serum sample a t time of amputation
of chondrosarcoma; n o metastasis
Reticulum cell sarcoma present 1 yr
post-diagnosis; no metastasis
Serum sample a t time of amputation of
rhabdomyosarcoma; metastasis present

Therapy

SCL
(rg/100 ml)

SZL
(pg/lOO ml)

5000R T o
1.5 yr prior

179

None

155

76

1200R T o t o
lungs 4 mo prior
5000R @ T oto
abdomen
14 mo prior
None

196

65

223

96

200

87

Transfer factor

104

123

None

216

127

None

169

325

None

171

104

None

142

278

None

165

148

Transfer factor

135

129

Transfer factor

152

100

2000R T o
2 mo prior
Same

172

112

164

180

None

149

82

None

165

96

7000R W o
1 yr prior
Transfer factor

127

78

97

160

Transfer factor

182

110

Transfer factor

196

86

None

156

68

None

174

106

None

168

108

None

140

198

77

The values for patients 7-11, rising the initial value for 10 and the second value for 11, comprised the primary
osteosarcoma group. Patients 1-6, using the initial value for 5, comprised the metastatic osteosarcoma group.
The amputated osteosarcoma; no demonstrable tumor group was computed from the second values for patients
5 and 14 and the average of the last two values for patient 10. T h e other primary sarcomas without metastasis
group was computed from the initial value for patient 11 and from patients 12, 17, and 18.

362
CANCER
January 1976
VOl. 37
category (Table 3) since she was in the early stages while depressed zinc and elevated copper
of pregnancy when both samples were drawn. levels were associated with pulmonary metasHowever, the pregnancy was electively terminated tases. Patients whose primary osteosarcomas
a t the 5th month, and follow-up serum samples will
were removed and who were clinically free of
be obtained. She is clinically tumor-free, and both
disease were found t o have serum copper and
serum samples were taken 12-18 months postzinc levels which approached normal values
amputation.
and were lower than patients with primary
0s
teosarcomas.
DISCUSSION
Tessmer et a1.22showed that a rise in serum
T h e increased serum copper levels observed copper level has a positive correlation to inin patients with osteosarcoma and other sar- creased numbers of bone marrow blast cells
in leukemic patients. Delves et aL4 showed a
comas appear to be positively correlated with
the extent and activity of the disease. Most similar correlation of plasma copper:zinc
importantly, we found that patients with pul- ratios in bone marrow aspirates of leukemic
monary metastases from osteosarcoma had children. Mac Millan et al.I4 have observed an
greater elevations in serum copper level than increase in both serum copper and zinc levels
did patients with only primary tumors; in that appears to correlate positively with the
contrast, the zinc level was lower in patients number of peripheral blast cells in a dog with
with metastases than in those with primary radiation-induced myeloproliferative disease
tumors only. Thus, not only may elevated characterized by significant leukemic infiltration of liver, spleen, lymph nodes, lung, and
serum copper be an aid in the differential
diagnosis of sarcoma, but the ratio of copper heart. O u r findings on SCL elevations in osteoto zinc in patients with osteosarcoma appears sarcoma suggest that the observed correlations
to be an indicator of the presence of pul- of SCL and SZL with blast cells in leukemic
monary metastasis. Furthermore, the three patients may not be a primary relationship,
patients with the highest SCLs or SCL:SZL but an indicator of the progress and state of
the disease.
ratios had the poorest survival, all dying of
Hrgovcic et a1.lQhave stated that the mechaextensive pulmonary metastases within 3
months of the time of serum analysis. I t ap- nism of SCL elevation is not known, although
pears that the determination of serum copper it has been shown that 9GYo of the copper in
and zinc levels in osteosarcoma patients should normal serum is bound to the enzyme cerulobe of value in prognosis and evaluation of plasmin.9 Shifrine and Fisher20 have found in
therapy. As reviewed in the introduction, a preliminary study that osteosarcoma patients
other investigators have reported on the value with elevated serum copper levels also have eleof serum copper level determinations in the vated serum concentrations of ceruloplasmin.
Although, as reviewed by Hrgovcic,lo hyperevaluation of Hodgkins disease,23124 leukemia,4,10 and malignant 1yrnphoma.ll Delves cupremia is associated with a variety of
et al.4 have reported that plasma copper:zinc abnormal physiologic states including hematoratios are elevated in untreated leukemic chil- poietic neoplasias, pregnancy, chemical condren, and that the ratio appears to be useful traception, and acute and chronic infections,
in evaluating the patients response to therapy. the determination of SCL and SZL appears to
In osteosarcoma, the SCL:SZL ratio was found be a useful clinical test in the differential diagto be very useful; elevated zinc and copper nosis, prognosis, and therapy evaluation of
levels were associated with primary tumors human osteosarcoma.

REFERENCES
1. Addink, N. W. H., and Frank, L. J. P.: Remarks apropos of analysis of trace elements i n human
tissue. Cancer 12:544-551, 1959.
2. Cancer Facts and Figures. American Cancer Society, 1974.
3. Chahovitch, X.: GI. srpske Akad. Nankz. Od.
nied. Naukn., Vol. 215, p. 143, 19855. I n Progress in

Experimental Tumor Research, Vol. 12, Basel, Switzerland, S. Karger, A.G., 1969.
4. Delves, H. T., Alexander, F. W., and Lay, H.:
Copper and zinc concentration in the plasma of leukemic children. BT. J . Haematol. 24:525-531, 1973.
5. DeWys, W., Pories, W. J., Richter, M. C., and
Strain, W. H.: Inhibition of Walker 256 arcinmar-

No. 1

SERUM
COPPERAND ZINCI N SARCOMA
PATIENTS

coma growth by dietary zinc deficiency. Proc. Sac. Ex@.

Ili01. Aled. 135:17-22, 1970.

6 . Diem. K., and Lentner, C., Eds.: Scientific Tables.
B a d , Switzerland, CIBA-Geigy Limited, 1971; p. 809.
7. Fisher, G. L.: Trace element changes in radiationrel;ltetl and spontaneous disease states: a preliminary
report. I n Annual Report, Univ. of Calif., Davis, CA,
Radiobiology Laboratory, UCD 472-121, 1974; pp.
76-79.
8. Friedman, M. A., and Carter, S. K.: T h e therapy
of ostcogenic sarcoma. J . 5ut.g. Oncol. 4:48?-510, 1972.
9. Gubler, C. I., Lahey, M. E., Cartwright, C. E.,
antl Wintrobe, M. hf.: Studies on copper metabolism.
IX. T h e transportation of copper in blood. J. Biol.
C h e / I l . 224:405-413, 1957.
10. Hrgobcic, M., Tessmer, C. F., Brown, B. W.,
Wilbur, J. R., Mumford, D. M., Thomas, F. B.,
Scliullenberger, C. C., and Taylor, G.: Serum copper
studies in the lymphomas and acute leukemias. Frog.
Clin. Cnncer 5:121-153, 1973.
11. Illicin, G.: Serum copper and magnesium levels
i n leukaemia and malignant lymphoma. Lancet 21036,
1971.
12. Jaffe, N., Kim, B. S., antl Vanter, G. F.: Hypocalcemia-a complication of childhood leukemia. Cantor 29:392-398, 1972.
13. Janes. J. M., McCall, J. T., and Elveback, L. R.:
Trace metals in human osteogenic sarcoma. hfayo
Clinic Proc. 47:4i6-478, 1972.
14. Mac Millan, K. L., Wilson, F. D., Fisher, G. L.,
and Chrisp, C. E.: Status of &Copilot study. In Annual
Report, Univ. of Calif., Davis, CA, Radiobiology Laboratory, UCD 472-121, 1974; pp. 13-17.

Fisher et al.

363

15. Mikac-Devic, D.: Methodology of zinc determinations and the role of zinc in biocheniical processes.
A d v . C l i n . Chem. 13271, 1970.
16. Moe, P. J.: Serum iron as differential diagnostic
aid in acute leukemia in children. Scand. J. Haeniatol.
6:270-273, 1969.
17. Remington, R. D., and Schork, M. A.: Statistics
with Applications to the Biological and Health Sciences.
Englewood CliRs, Prentice-Hall, Inc., 1970.
18. Rubin, H., and Kordi, T.: Stimulation of DNA
synthesis and 2-deoxy-D-glucose transport in chick
embryo cultures by excessive metal concentrations antl
by a carcinogenic hydrocarbon. J . Cell Physiol. 81:
387-396, 1973.
19. Schajowicz, F.: Giant cell tumors of bone (osteoclastoma). I. Bone Joint S w g . 43:l-29, 1961.
20. Shifrine, M., and Fisher, G. L.: Ceruloplasm
in levels in sera from human patients with osteosarcoma. Cnncer (in press).
21. Szmigielski, S . , and Litwin, J.: T h e histochemical
demonstration of zinc in blood granulocytes-the
new
test in diagnosis of neoplastic diseases. Cancer 1:13811384, 1964.
22. Tessmer, C. F.. Hrgovcic, M., Brown, B., Wilbur, J., and Thomas, F.: Serum copper correlations
with hone marrow. Cancer 29:173-179, 1972.
23. Tessmer, C. F., Hrgovcic, M., and Wilbur, J.:
Serum copper in Hodgkins disease in children. Cancer
31 ~303-315, 1973.
24. Warren, R. L., Jelliffe, A. M., Watson, J. V.,
and Hobbs, C. B.: Prolonged observations on variations
in the serum copper in Hodgkins disease. Clin. Radiol.
20~247-256, 1969.